Construction, expression, and identification of the gene of human anti-prostate specific membrane antigen single-chain antibody / 中华男科学杂志
National Journal of Andrology
;
(12): 1063-1067, 2014.
Article
Dans Chinois
| WPRIM
| ID: wpr-319567
ABSTRACT
<p><b>OBJECTIVE</b>To construct, express and purify human fusion proteins composed of a single-chain antibody fragment scFv that recognizes the prostate specific membrane antigen (PSMA) protein, Fdt, HA2 and tp, and to analyze the binding activity of the expressed fusion proteins.</p><p><b>METHODS</b>The fusion protein genes scFv, scFv-tp, and scFv-Fdt-HA2-tp were amplified by PCR, and the genes obtained were then cloned into the expression vector pET28 and expressed in E. coli BL21. The expressed products were identified by SDS-PAGE and Western blot and purified with Ni(2+)-NTA chelating agarose. The antigen-binding activity of the fusion proteins was determined by ELISA.</p><p><b>RESULTS</b>The human anti-PSMA fusion gene was successfully constructed and expressed in M15 as the inclusion body after induced with IPTG. All the target proteins expressed could bind the PSMA antigen.</p><p><b>CONCLUSION</b>Fusion proteins can specifically bind the PSMA antigen. This finding contributes to the study of the targeted delivery of siRNA.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Protéines de fusion recombinantes
/
Réaction de polymérisation en chaîne
/
Clonage moléculaire
/
Petit ARN interférent
/
Glutamate carboxypeptidase II
/
Électrophorèse sur gel de polyacrylamide
/
Allergie et immunologie
/
Escherichia coli
/
Anticorps à chaîne unique
/
Génétique
Type d'étude:
Étude pronostique
Limites du sujet:
Humains
/
Mâle
langue:
Chinois
Texte intégral:
National Journal of Andrology
Année:
2014
Type:
Article
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